Freeze-dried antibacterial tablet and preparation method thereof

Abstract

According to the freeze-dried antibacterial tablet disclosed by the invention, nano-silver and lysozyme are compounded, and the antibacterial effect is remarkably improved by utilizing the synergistic effect of the nano-silver and the lysozyme. The antibacterial effect is improved by compounding the lysozyme, and the potential risk to the human body and the environment is reduced under the condition that the concentration of the nano-silver is not increased. The problems that in the prior art, a single antibacterial agent is limited in effect and poor in safety are solved, and the efficient, safe and lasting sterilization effect is achieved. The auxiliary materials such as carbomer and polyethylene glycol are added, so that safety and effectiveness are achieved, and the experience feeling is good; the production process is more advanced freeze-drying production process at present; the maximum retention of the effective components is ensured. And a propeller is arranged, so that the use is convenient, and the bacteriostatic tablet can directly act on an affected part to play a bacteriostatic effect.

Description

Technical Field

[0001] The invention belongs to the field of biological nanomaterials and sterilization, and specifically relates to a freeze-dried antibacterial tablet and a preparation method thereof. Background Art

[0002] In the field of modern medicine and public health, the application of antimicrobial and antiseptic agents is crucial, as bacterial and viral infections are the main causes of many diseases. Therefore, the development of effective antimicrobial technologies is of great significance. Silver ions (Ag+) and lysozyme are two substances with significant antimicrobial properties.

[0003] The chemical structure of silver gives it high catalytic ability and oxidizing properties. Silver ions can destroy proteases in bacterial cells, inhibit their activity, and cause bacterial death. Silver ions can also attract sulfhydryl groups (-SH) in bacteria, quickly bind to and inactivate bacterial proteases, and produce atomic oxygen (O) with strong oxidizing properties to directly kill bacteria. In addition, silver ions can also coagulate viral protein molecules and bind electron donors on their DNA molecules, resulting in viral inactivation. Studies have shown that 0.01ppm of silver ions in water can completely kill E. coli in water and prevent new bacterial colonies from growing within 90 days. Existing silver ion antibacterial technology has been widely used in various antibacterial products, such as silver ion coatings, silver ion water treatment systems, and silver ion medical dressings. These products use the strong oxidizing properties and protease inactivation of silver ions to achieve effective antibacterial effects. However, these applications usually require strict control of silver ion concentrations to avoid potential harm to humans and the environment.

[0004] Lysozyme is a naturally occurring antibacterial enzyme with a molecular weight of 14.4 kDa. It is part of the innate immune system and destroys the bacterial cell wall by catalyzing the hydrolysis of the 1,4-β-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine residues in the peptidoglycan of the bacterial cell wall, leading to bacterial lysis and death. Lysozyme can destroy the bacterial cell wall, allowing it to exert a significant antibacterial effect even at low concentrations, and is therefore considered a safe and effective natural antibacterial agent. However, the use of lysozyme alone has limited effect on some Gram-negative bacteria and drug-resistant bacteria.

[0005] Patent CN117204439A discloses a novel enzyme-silver antiviral and antibacterial composite liquid preparation, which can be used for air disinfection, article disinfection, etc.; Patent CN115154594A discloses a lysozyme-silver nitrate complex freeze-dried powder, which is prepared by reaction under visible light irradiation, and can also be used in the fields of medical equipment, disinfection products, and cosmetics. The bactericides disclosed in the above two patents are in liquid dosage form when used. However, the female antibacterial agent dosage forms on the market are mostly antibacterial gels or lotions; because the physiological structure of women is different from that of men, there are many changes during use, and the gel cannot accurately control the amount during use, and the lotion needs to be diluted with water in a certain proportion, and then the diluted lotion is used to clean the affected area. The lotion can only clean the surface of the vagina and cannot really enter the affected area, so it cannot achieve the ideal antibacterial effect; however, for women who travel or go on business trips: first, it is inconvenient to carry; second, it is inconvenient to use and cannot achieve the ideal antibacterial effect. Summary of the invention

[0006] Purpose of the invention: The purpose of the present invention is to provide a female antibacterial agent, especially a lyophilized agent containing lysozyme and nanosilver. Through the compounding of nanosilver and lysozyme, the limited effect and safety problems of a single antibacterial agent in the prior art are solved, and a high-efficiency, safe and long-lasting bactericidal effect is achieved. Especially as a female antibacterial agent, it is convenient for women to carry and use.

[0007] Technical solution: The freeze-dried antibacterial tablet described in the present invention comprises lysozyme, nanosilver, an adhesive and a thickener.

[0008] Furthermore, the adhesive is polyethylene glycol 8000 (PEG8000).

[0009] Furthermore, the thickener is a carbomer copolymer.

[0010] The present invention also provides a method for preparing the above-mentioned freeze-dried antibacterial tablet: 1) mixing lysozyme, nanosilver, a binder and a thickener in a certain proportion and dissolving them in water and stirring them evenly to form a mixed aqueous solution; 2) Use an alkaline regulator to adjust the pH value of the solution in step 1) to 6.2-8.2; 3) dripping the solution obtained in step 2) into a sheet mold, and placing the mold in a freezer to freeze; 4) The antibacterial tablet obtained in step 3) is transferred to a vial and freeze-dried in a vacuum freeze dryer to obtain a freeze-dried antibacterial tablet.

[0011] Furthermore, in step 1), the mass percentage of lysozyme in the mixed aqueous solution is 0.1-1.0wt%, and the content of nanosilver is 1ppm.

[0012] Furthermore, the mass percentage of the adhesive is 4-5wt%, and the mass percentage of the thickener is 0.10-0.15wt%.

[0013] Furthermore, the alkaline regulator in step 2) is sodium hydroxide.

[0014] Furthermore, in step 3), the temperature of the freezer is -20°C, the freezing time is 2h-5h, and the freeze-drying time in step 4) is 20-30h.

[0015] The present invention also provides the use of the freeze-dried antibacterial tablet in the preparation of female antibacterial drugs.

[0016] Beneficial effects:

[0017] The present invention compounds nanosilver and lysozyme, and utilizes the synergistic effect of the two to significantly improve the antibacterial effect. The antibacterial effect is improved by compounding lysozyme, and the potential risks to the human body and the environment are reduced without increasing the concentration of nanosilver. The limited effect and safety problems of a single antibacterial agent in the prior art are solved, and an efficient, safe and lasting bactericidal effect is achieved. The formula of this invention uses nanosilver and lysozyme as the components of the antibacterial tablet; after 2 minutes of action, the antibacterial rate of Candida albicans, Staphylococcus aureus and Escherichia coli reaches more than 90%, and has a strong antibacterial effect; with the addition of auxiliary materials such as carbomer and polyethylene glycol, it is safe and effective, and the experience is good; the production process uses the more advanced freeze-drying production process at present; ensuring the maximum retention of the active ingredients. The dosage is more precise, equipped with a propeller, and easy to use. The antibacterial tablet can act directly on the affected area to exert an antibacterial effect.

[0018] The freeze-dried product of the present invention is light in weight and small in size, and can be stored at room temperature and transported at room temperature; however, the storage temperature of liquid preparations is strictly required, and they need to be stored and transported at 2-8°C; in addition, the product is easily redissolved by adding water, and the freeze-dried solid material presents a porous structure due to the sublimation of tiny ice crystals, and maintains the original volume when frozen, so it is very easy to dissolve and recover after adding water; after freeze-drying, the product is always in a vacuum state, which can prevent oxidation and has a longer shelf life. DETAILED DESCRIPTION

[0019] In order to deepen the understanding of the present invention, the present invention will be further described in detail below in conjunction with examples. The examples are only used to explain the present invention and do not constitute a limitation on the protection scope of the present invention.

[0020] The names, models and manufacturers of the raw materials used in the examples are as follows: PEG8000 P8260 Beijing Soleboc Co., Ltd.; Carbomer copolymer type B Guangzhou Kangqiao Hamp Pharmaceutical Co., Ltd.; Lysozyme Hubei Kemaidi Chemical Co., Ltd.; Nanosilver solution AGS-WMB300C Shanghai Huzheng Industrial Co., Ltd.

[0021] Example 1 Synergistic effect experiment of lysozyme and nanosilver solution Prepare antibacterial aqueous solutions with different contents, the specific content ratios are as follows: Group 1: 5ppm nanosilver; Group 2: 1ppm nanosilver; Group 3: 1ppm nanosilver, 0.1% lysozyme; Group 4: 1ppm nanosilver, 0.3% lysozyme; Group 5: 1ppm nanosilver, 0.5% lysozyme; Group 6: 1ppm nanosilver, 0.7% lysozyme; Group 7: 1ppm nanosilver, 1% lysozyme; Group 8: 1% lysozyme.

[0022] Test method: Take 0.1mL of Candida albicans ATCC10231, 0.1mL of Staphylococcus aureus ATCC6538, and 0.1mL of Escherichia coli 8099 respectively, add them to sterile test tubes containing 10mL of LB liquid culture medium, and then add the prepared 8 groups of test sample solutions to the aforementioned test tubes. At the same time, set up a blank control (only LB liquid culture medium and bacterial solution are added to the test tube).

[0023] Cultivate at 37℃, measure the OD600nm of each test tube bacterial solution after 24 hours (adjust to zero with LB culture medium), and record and calculate the antibacterial rate of each product at different concentrations. As shown in Table 1, the experimental results show that the fifth group: 1ppm nanosilver, 0.5% lysozyme has a higher antibacterial rate against the three bacterial solutions than other experimental groups, and has a strong antibacterial effect.

[0024] Antibacterial rate = (A-blank - A-sample) / A-blank * 100% Note: A: 24h OD value of blank control group; blank control has no antibacterial effect, is bacterial culture solution, and the OD value is 0.8.

[0025] Sample A: 24h OD value of the sample group.

[0026] Table 1 Antibacterial rate of each group of antibacterial aqueous solution

[0027] Example 2 Preparation method of freeze-dried antibacterial tablets 1) lysozyme, nanosilver, PEG8000 and carbomer copolymer are mixed and dissolved in water according to a certain proportion and stirred to form a mixed aqueous solution; finally, the volume is fixed to 100 ml with the aqueous solution; the final solution contains 5% PEG8000, 0.12% carbomer, 0.5% lysozyme, and 1 ppm nanosilver; 2) Use an alkaline regulator to adjust the pH value of the solution in step 1) to 6.2-8.2; after fully stirring for 10 minutes again, take part of the semi-finished working solution and perform eutectic point detection; 3) dripping the solution obtained in step 2) into the sheet mold using an automatic drip irrigation machine, and transferring the mold to a -20°C freezer for freezing for more than 2 hours until the solution is fully frozen and formed; 4) The antibacterial tablet obtained in step 3) is transferred to a vial and freeze-dried in a vacuum freeze dryer for about 24 hours to obtain a freeze-dried antibacterial tablet; the freeze-drying procedure of step 4) is shown in Table 2 below.

[0028] Table 2 Freeze-drying procedure

[0029] Example 3 Preparation method of freeze-dried antibacterial tablets The difference from Example 2 is that the solution obtained in step 1) contains 4% PEG8000, 0.12% carbomer, 0.5% lysozyme, and 1 ppm nanosilver.

[0030] Example 4 Preparation method of freeze-dried antibacterial tablets The difference from Example 2 is that the solution obtained in step 1) contains 4% PEG8000, 0.15% carbomer, 0.5% lysozyme, and 1 ppm nanosilver.

[0031] Example 5 Preparation method of freeze-dried antibacterial tablets The difference from Example 2 is that the solution obtained in step 1) contains 5% PEG8000, 0.15% carbomer, 0.5% lysozyme, and 1 ppm nanosilver.

[0032] Example 6 Appearance powder loss test and loss rate calculation According to the experimental regulations: the loss rate of the antibacterial tablet appearance powder loss test should be less than 8%; the test steps are: weigh the 4 groups of freeze-dried products before transportation shaking test: Example 2: 22.3 mg, Example 3: 21.5 mg, Example 4: 22.6 mg, Example 5: 23.3 mg, put them in a rotary mixer, fix the position, adjust the speed knob to 40RPM, and time for 1 hour; weigh the antibacterial tablets after the test according to different speeds, and calculate the loss rate (weight before test-weight after test) / total weight before test*100%; see the following Table 3 for details: Table 3

[0033] The experimental data show that the product obtained in Example 5 is more in line with the formulation process of freeze-dried antibacterial tablets.

[0034] Example 7 Skin and vaginal mucosal irritation test The present invention is an antibacterial tablet. From the perspective of female hygiene products, the antibacterial tablet can be directly used on the vaginal mucosa or diluted and used on intact skin for antibacterial purposes; it is convenient to use and carry.

[0035] The freeze-dried antibacterial tablets prepared in Example 5 were used for testing, and intact skin and vaginal mucosal irritation tests were performed according to method 2.3.3 of the 2002 edition of the Technical Specifications for Disinfection; local reactions on the skin and mucosa were observed, and the irritation reactions were scored.

[0036]

[0037] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A freeze-dried antibacterial tablet, characterized in that: Contains lysozyme, nanosilver, binder and thickener.

2. The freeze-dried antibacterial tablet according to claim 1, characterized in that The binder is polyethylene glycol 8000.

3. The freeze-dried antibacterial tablet according to claim 1, characterized in that The thickener is a carbomer copolymer.

4. The freeze-dried antibacterial tablet according to any one of claims 1 to 3, characterized in that: The preparation method is as follows: 1) mixing lysozyme, nanosilver, a binder and a thickener in a certain proportion and dissolving them in water and stirring them evenly to form a mixed aqueous solution; 2) Use an alkaline regulator to adjust the pH value of the solution in step 1) to 6.2-8.2; 3) dripping the solution obtained in step 2) into a sheet mold, and placing the mold in a freezer to freeze; 4) The antibacterial tablet obtained in step 3) is transferred to a vial and freeze-dried in a vacuum freeze dryer to obtain a freeze-dried antibacterial tablet.

5. The freeze-dried antibacterial tablet according to claim 4, characterized in that: Step 1) The mass percentage of lysozyme in the mixed aqueous solution is 0.1-1.0wt%, and the content of nanosilver is 1ppm.

6. The freeze-dried antibacterial tablet according to claim 4, characterized in that: The mass percentage of the adhesive is 4-5wt%, and the mass percentage of the thickener is 0.10-0.15wt%.

7. The freeze-dried antibacterial tablet according to claim 4, characterized in that: The alkaline regulator in step 2) is sodium hydroxide.

8. The freeze-dried antibacterial tablet according to claim 4, characterized in that: In step 3), the temperature of the freezer is -20°C, the freezing time is 2h-5h, and the freeze-drying time in step 4) is 20-30h.

9. Use of any one of the freeze-dried antibacterial tablets according to claims 1 to 8 in the preparation of female antibacterial drugs.

Images